目的　克隆THANKcDNA ,并在大肠杆菌中进行表达。方法　采用RT PCR技术 ,从人外周血单个核细胞的总RNA中扩增人THANK全长编码区基因及THANK胞外区编码基因 ,PCR产物直接克隆于pMD 18T载体中 ,重组克隆进行DNA测序。将测序证实的THANK胞外区基因亚克隆到原核表达载体pET 11a中。阳性重组子 ,以 1mmol/LIPTG进行诱导表达 ,以SDS PAGE分析THANK胞外区的表达。对表达的蛋白作初步纯化处理后 ,进行生物学活性检测。结果　RT PCR扩增出一个85 8bp的DNA片段 ,限制性内切酶图谱分析和测序结果显示 ,该片段为编码人THANK的cDNA ,与公布的人THANK基因序列一致。将胞外区片段克隆入表达载体 ,转化大肠杆菌表达后发现 ,与阴性对照相比 ,在相对分子质量 (Mr) 2 6× 10 4 处多显示出一条条带。对该蛋白进行初步活性测定显示其可显著地抑制U937细胞的生长。结论　本实验成功地克隆了人THANK基因 ,并将其可溶性胞外区片段在大肠杆菌中进行了表达 ,表达的重组蛋白可抑制U937细胞的生长。这为进一步进行THANK基因的功能研究及其开发和临床应用奠定了基础 Objective To clone THANK cDNA and express it in E. coli. Methods The full-length human THANK coding region gene and THANK extracellular region gene were amplified by RT-PCR from total RNA of human peripheral blood mononuclear cells. The PCR products were directly cloned into pMD 18T vector and cloned into DNA for sequencing. The sequencing confirmed THANK extracellular region gene was subcloned into the prokaryotic expression vector pET11a. Positive recombinant was induced by 1 mmol / L IPTG, and the expression of THANK extracellular domain was analyzed by SDS PAGE. The expressed protein for the initial purification treatment, the biological activity test. Results A 858bp DNA fragment was amplified by RT-PCR. The results of restriction enzyme analysis and sequencing showed that the fragment was cDNA encoding human THANK, which was consistent with the published human THANK gene sequence. The fragment of extracellular region was cloned into expression vector and transformed into Escherichia coli to express a band more than 26 × 10 4 in molecular weight (Mr) compared with negative control. The preliminary activity of the protein showed that it can significantly inhibit the growth of U937 cells. Conclusion The human THANK gene was successfully cloned and the soluble extracellular region of the gene was expressed in E. coli. The expressed recombinant protein inhibited the growth of U937 cells. This laid the foundation for the further study on the function of THANK gene and its development and clinical application